Abstract
The Escherichia coli siderophore enterobactin is assembled from 2,3-dihydroxybenzoate (2,3-DHB) and l-serine by the nonribosomal peptide synthetases EntB and EntF. The processive thiol-template strategy used can be sabotaged by EntB misacylation. Through in vitro kinetic analysis, we demonstrate two potential routes to EntB misacylation and provide evidence for two mechanisms by which the hot dog-fold thioesterase EntH can potentially prevent or reverse EntB misacylation.
Publication types
-
Research Support, N.I.H., Extramural
MeSH terms
-
Acyl Coenzyme A / metabolism
-
Benzophenones / metabolism
-
Catalysis
-
Enterobactin / biosynthesis*
-
Enterobactin / chemistry
-
Escherichia coli / enzymology*
-
Escherichia coli Proteins / chemistry*
-
Escherichia coli Proteins / metabolism*
-
Holoenzymes / metabolism
-
Hydrogen-Ion Concentration
-
Hydrolysis
-
Kinetics
-
Protein Structure, Secondary
-
Spectrometry, Mass, Electrospray Ionization
-
Substrate Specificity
-
Temperature
-
Thiolester Hydrolases / chemistry*
-
Thiolester Hydrolases / metabolism*
-
Time Factors
Substances
-
Acyl Coenzyme A
-
Benzophenones
-
Escherichia coli Proteins
-
Holoenzymes
-
Enterobactin
-
EntH protein, E coli
-
Thiolester Hydrolases
-
2,4-dihydroxybenzophenone